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WO2024227514A1 - Déphosphorylation de lait écrémé, de lait ultra-filtré ou d'isolat de caséine micellaire - Google Patents

Déphosphorylation de lait écrémé, de lait ultra-filtré ou d'isolat de caséine micellaire Download PDF

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Publication number
WO2024227514A1
WO2024227514A1 PCT/EP2023/061832 EP2023061832W WO2024227514A1 WO 2024227514 A1 WO2024227514 A1 WO 2024227514A1 EP 2023061832 W EP2023061832 W EP 2023061832W WO 2024227514 A1 WO2024227514 A1 WO 2024227514A1
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Prior art keywords
milk
protein
mci
skim milk
kcal
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PCT/EP2023/061832
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English (en)
Inventor
Hilde Ruis
Liya YI
Original Assignee
N.V. Nutricia
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Publication date
Application filed by N.V. Nutricia filed Critical N.V. Nutricia
Priority to PCT/EP2023/061832 priority Critical patent/WO2024227514A1/fr
Priority to PCT/EP2024/062051 priority patent/WO2024227851A1/fr
Publication of WO2024227514A1 publication Critical patent/WO2024227514A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/15—Reconstituted or recombined milk products containing neither non-milk fat nor non-milk proteins
    • A23C9/1512—Reconstituted or recombined milk products containing neither non-milk fat nor non-milk proteins containing isolated milk or whey proteins, caseinates or cheese; Enrichment of milk products with milk proteins in isolated or concentrated form, e.g. ultrafiltration retentate
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C7/00—Other dairy technology
    • A23C7/04—Removing unwanted substances other than lactose or milk proteins from milk
    • A23C7/043—Removing unwanted substances other than lactose or milk proteins from milk using chemicals in liquid or solid state, e.g. flocculating, adsorbing or extracting agents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
    • A23J1/20—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
    • A23J1/20—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey
    • A23J1/202—Casein or caseinates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/04—Animal proteins
    • A23J3/08—Dairy proteins
    • A23J3/10—Casein
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/325—Working-up of proteins for foodstuffs by hydrolysis using chemical agents of casein
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/19—Dairy proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C19/00—Cheese; Cheese preparations; Making thereof
    • A23C19/02—Making cheese curd
    • A23C19/05—Treating milk before coagulation; Separating whey from curd
    • A23C19/052—Acidifying only by chemical or physical means

Definitions

  • the present invention is in the field of a high-protein liquid enteral nutritional composition containing predominant amounts of micellar casein. More particularly, the present invention relates to dephosphorylation of skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI). More in particular, the present invention addresses the problem of reducing phosphorus content of skim milk, ultra-filtered milk (UF milk, including MPC or MPI) or micellar casein isolate (MCI) and to a liquid shelf-stable and heat-sterilized enteral nutritional composition comprising 16 - 35 en% protein and 2.0 - 3.0 kcal/ml caloric content with a phosphorus content compliant with the requirements of foods for special medical purposes (FSMP).
  • FSMP special medical purposes
  • the present invention is in the field of high-protein liquid enteral nutritional compositions which are ready-to-use compositions with low intake volume, and enriched with all essential minerals, vitamins and trace elements.
  • Use of high-protein nutritional supplement lowers the mortality and complication rates of malnourished patients, for which patients it is key to provide the required nutritional support and calories that they will otherwise not achieve from normal eating behaviour.
  • using high-protein nutritional supplement provides the opportunity to improve the quality of life of malnourished patients and/or elderly.
  • micellar casein products are MCI88 from Friesland Campina, with phosphorous levels of 1500-1700 mg/100 g powder (and with 85g protein/100 g) i.e. 17.6 - 20 mg phosphorous per g protein.
  • Casein provides about half of the P of milk (22% organic, 32% inorganic), there being about 1 g P per kg milk [source: Walstra, P., Wouters, J. T., & Geurts, TJ. (2006). Part 2: Processes, in Dairy science and technology. CRC press: Boc Raton, USA p. 207-272], Casein contains two types of phosphorus, i.e. organic and inorganic phosphorous. P is part of colloidal calcium phosphate (inorganic-P) or covalently bound to caseins as phosphate groups (casein-P). Casein micelles typically contain both organic (46%) and inorganic phosphorus (64%).
  • micellar casein products Phosphorous removal from such micellar casein products is a delicate matter, since together with the calcium the phosphorous is a key factor in stabilizing the micellar structure, while it is the micellar structure that is key to keep high-protein compositions stable.
  • the core of a casein micelle is bound by calcium phosphate nanoclusters (CCP or casein phosphopeptide), and these CCP nanoclusters are important for maintaining the casein micelle structures heat-stable. Without CCP the micelles would fall apart. While phosphorous can be washed out, in order to maintain product stability, and simultaneously retain increased protein concentrations yet reduced viscosities, it is important to keep the integrity of the micellar casein essentially intact.
  • high-protein liquid shelf-stable enteral nutritional compositions commercially available typically contain total phosphorus levels up to 300 mg/100 mL, which is much higher than the maximal level of 192 mg/100 ml (or 30 - 80 mg/lOOkcal) required by FSMP.
  • the major part of total phosphorus (about 250 mg/100 mL) is from the micellar casein source, and only a minor part of total phosphorus (about 50 mg/100 mL) originates from K2PO4.
  • at least 23% P should be reduced, and clearly such numbers can only be achieved when making use of MCI which exhibits reduced P levels.
  • W02009/072885 discloses a problem associated with the use of micellar casein in the production of a high protein liquid enteral nutritional composition and further containing acids, in particular citric acid, is the formation of calcium-acid complexes, such as calcium citrate.
  • acids in particular citric acid
  • a certain Ca-ion activity is beneficial to maintain a desired viscosity of the composition during processing of the composition, e.g. during pasteurisation and/or sterilisation.
  • the viscosity of the composition during processing including heat-sterilization is controlled by using a mixture of micellar casein and caseinate.
  • WO2013/129925 discloses a liquid enteral nutritional composition with a high energy content comprising 6 to 20 g / 100 ml protein, said protein comprising micellar casein and lactic acid.
  • the purpose of this application to reduce viscosity of a protein-dense composition with increased levels of micellar casein by adding lactic acid rather than using citric acid.
  • micellar casein While high amounts and concentrations of micellar casein are preferred to produce high-protein liquids, the disadvantage is that this yields an increase in phosphorous concentrations which compromises on FSMP requirements, and where it is challenging to remove the phosphorous without destabilizing the micelles. Therefore, there is a need in the art to reduce the phosphorus content of high-protein liquid enteral nutritional composition which are high in micellar casein, ultimately to comply with the FSMP requirements.
  • the phosphorous content should be below 15mg, preferably below 14 mg even more preferably below 13.3 mg phosphorous per gram protein.
  • micellar casein structure In essence, the lower the better without compromising the micellar casein structure; it is only with those restrictions that high-protein compositions with protein levels beyond 12 g/100 ml up to 14.4 g/100 ml compliant with FSMP requirements on P could be produced (using Nitrogen factor 6.25).
  • the present invention provides a process to reduce the phosphorus content of skim milk, ultrafiltered milk (UF milk), including milk protein concentrate (MPC) and milk protein isolate (MPI), or micellar casein isolate (MCI), and which dephosphorylated product makes a very suitable ingredient to produce a high-protein liquid shelf-stable (heat-sterilized) enteral nutritional composition that complies with the FSMP requirements, particularly with regard to phosphorous content (a maximum of 192 mg/100 ml or 30 - 80 mg/lOOkcal according to FSMP directive 1999/21/EC).
  • Phosphorous levels in skim milk, UF milk and MCI as commercially marketed are about 1700 mg P per 100 g powder, or 20 mg P per g protein, which is too high to implement higher amounts of these proteins in high-protein products.
  • acidification to pH 6.0, cooling to 4 °C and using 70 mEq. gluconate the dephosphorylation method of the invention makes it possible to reduce to 75 mg P per 100 g powder or 12.3 mg P per g protein, which was a reduction of 38.3%.
  • the gluconate or maleate chelators manage to bind to the calcium and make it possible to wash out phosphorous while stabilising a micellar casein structure.
  • the effects of the selective acidification and specific chelators is demonstrated in the examples, and it turned out that the ability to retain the heat-stable micellar structure yet reduce P was limited to gluconate and maleate, with the best results obtained with gluconate. Care is taken to control pH in order to minimize or even avoid acid gel formation of the micelles. As said, particularly good heat-stabilizing effects have been obtained with gluconate and maleate, particularly gluconate.
  • Citrate is a common calcium chelator in the field, but as it appeared it was not able to keep the micelles heat-stable, and the amount of phosphorous reduction was also limited. Acetate showed disappointing results presumably due to the low affinity for CCP, no phosphorous reduction was observed. Similar results are expected with lactate, which - compared to acetate - has even worse calcium chelating affinity.
  • lactate which - compared to acetate - has even worse calcium chelating affinity.
  • citrate is believed to lead to CPP completely dissolve from the caseins, and, as explained here above, without CCP these caseins fall apart.
  • lactate and acetate show no affinity for CCP.
  • the present invention provides a process for dephosphorylation of skim milk, ultra-filtered milk (UF milk), including MPC and MPI, or MCI comprising;
  • step (iv) washing the skim milk, UF milk or MCI (of step (iii)) to remove phosphorus, thus dephosphorylating the skim milk, UF milk or MCI.
  • the total phosphorous content of the starting material i.e. skim milk, UF milk or MCI
  • the amount of total phosphorus in commercially available MCI is about 1500 - 1700 mg per 100 g, and with about 85/100 g protein, the amount of P in such MCI powders is typically about 18 - 20 mg per g protein.
  • micellar casein in the other starting materials may be lower than calculated here above for MCI, since the sole P source in these other milk sources is the micellar casein, the same 18 - 20 mg per g protein is also typical for those starting materials.
  • the invention pertains to dephosphorylated skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI) with gluconate and/or maleate, comprising 20 to 40 % less total phosphorus than the original skim milk, UF milk or MCI which has not been subjected to dephosphorylation according the invention (on total content).
  • the 'dephosphorylated' skim milk, UF milk (including MPC, MPI) or MCI preferably has a P content of less than 15.0 mg P per g protein, more preferably below 14.0 mg even more preferably below 13.3 mg phosphorous per gram protein. While there is no lower limit, in practice the amount of P is preferably above 10 mg phosphorous per gram protein.
  • dephosphorylated skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI) with gluconate and/or maleate is obtainable and preferably obtained by the process according to the invention and as described hereabove, and detailed throughout the application.
  • the present invention provides the use of dephosphorylated skim milk, UF milk or MCI for manufacturing a liquid, heat-sterilized, high-protein enteral nutritional composition comprising 2.0 - 3.0 kcal/ml wherein 16 - 35 en% is provided by protein, and wherein the protein comprises at least 70 wt% micellar casein, based on total protein.
  • the present invention provides a liquid heat-sterilized enteral nutritional composition
  • a liquid heat-sterilized enteral nutritional composition comprising 2.0 - 3.0 kcal/ml wherein 16 - 35 en% is provided by protein, wherein the protein comprises micellar casein (MC), whey protein (WP) and optionally caseinate (CAS), wherein there is at least 70 wt % MC and less than 15 wt % WP, based on total protein content, and wherein the composition has a total amount of phosphorous less than 192 mg/100 ml and/or 30 - 80 mg/lOOkcal, preferably at least 30 - 80 mg/100 kcal, most preferably less than 192 mg/100 ml and 30 - 80 mg/lOOkcal.
  • MC micellar casein
  • WP whey protein
  • CAS optionally caseinate
  • the composition is preferably at least characterized in terms of weight amounts per calories (mg/kcal), i.e. 30 - 80 mg/lOOkcal, more preferably 30 - 72 mg/100 kcal, even more preferably 30-64 mg/lOOkcal, in accordance with FSMP.
  • the composition further comprises gluconate and/or maleate. The presence of detectable amounts of gluconate and/or maleate in the composition is indicative of the use of dephosphorylated skim milk, UF milk or MCI, necessary to arrive at reduced amounts of phosphorous in compliance with FSMP.
  • the term 'high-protein' preferably means that the combination of caloric content and protein caloric content are selected such that there is 10 - 18 g/100 ml protein, preferably 12 - 18 g/100 ml protein in the composition.
  • the problems with compliance with FSMP plays particularly a part for these higher protein concentrations.
  • Figure 1 Effect of acidification on (a) stability (zeta-potential), (b) volume intensity (particle size) and (c) viscosity.
  • a process for dephosphorylation of skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI) comprising;
  • step (iv) washing the skim milk, UF milk or MCI, to remove phosphorus, thus dephosphorylating the skim milk, UF milk or MCI, preferably to an extent that the total phosphorous content of the skim milk, UF milk or MCI is reduced with at least 20%, preferably 20-40%, more preferably 30 - 40 % compared to the material provided to step (i).
  • gluconate and/or maleate chelator is sodium gluconate, potassium gluconate, disodium maleate and/or dipotassium maleate, preferably sodium gluconate and/or potassium gluconate.
  • a dephosphorylated skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI) obtainable by the process according to embodiment 1 or 2.
  • a dephosphorylated skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI) comprising (added) gluconate and/or maleate, the skim milk, UF milk of MCI having a total phosphorous content of less than 15.0 mg P per g protein, more preferably below 14.0 mg even more preferably below 13.3 mg total phosphorous per gram protein, wherein the dephosphorylated skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI) is preferably obtainable by the process according to embodiment 1 or 2.
  • dephosphorylated skim milk, UF milk or MCI for manufacturing a liquid, heat-sterilized high-protein enteral nutritional composition
  • a liquid, heat-sterilized high-protein enteral nutritional composition comprising 2.0 - 3.0 kcal/ml wherein 16 - 35 en% is provided by protein, and wherein the protein comprises at least 70 wt% micellar casein, based on total protein, wherein the composition has a total amount of phosphorous less than 192 mg/100 ml and/or 30 - 80 mg/lOOkcal, preferably at least 30 - 80 mg/100 kcal, most preferably less than 192 mg/100 ml and 30 - 80 mg/lOOkcal. 6.
  • a liquid heat-sterilized enteral nutritional composition comprising 2.0 - 3.0 kcal/ml wherein 16 - 35 en% is provided by protein, the combination of caloric content and relative protein caloric content selected such that there is 10 - 18 g/100 ml protein, preferably 12 - 18 g/100 ml protein in the composition, wherein the protein comprises micellar casein (MC), whey protein (WP) and optionally caseinate (CAS), wherein there is at least 70 wt % MC and less than 15 wt % WP, based on total protein content, and wherein the composition has a total amount of phosphorous less than 192 mg/100 ml and/or 30 - 80 mg/lOOkcal, preferably at least 30 - 80 mg/100 kcal, most preferably less than 192 mg/100 ml and 30 - 80 mg/lOOkcal.
  • MC micellar casein
  • WP whey protein
  • CAS optionally caseinate
  • liquid composition according to any one of embodiments 6 - 8, wherein the amount of protein is between 12 and 18 g/100 ml.
  • micellar casein to caseinate ranges from 90:10 to 60:40.
  • liquid composition according to any one of embodiments 6 - 10, wherein the amount of whey protein is less than 10 wt% based on total protein.
  • liquid composition according to any one of embodiments 6 - 11, wherein the composition comprises 75 mg P per 100 g dry weight, and/or 12.3 mg P per g protein.
  • liquid composition according to any one of embodiments 6 - 12, further comprising at least 2 mineral levels, more preferably at least 3, even more preferably at least 4, most preferably all mineral levels for Na, K, Cl, Ca and Mg within the ranges (mg/100 kcal) according to the table below:
  • the present invention provides a process for dephosphorylation of skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI).
  • skim milk, UF or MCI is acidified (i.e. pH ⁇ 7), but preferably not lower than 6.0. Below pH 6.0, acid formation of MCI was observed, and the desired micellar structure gets lost. In the example, this acid gel formation was observed at pH 5.8.
  • the starting material is preferably UF milk including MPC and MPI, or MCI.
  • Ultrafiltered milk is a subclassification of milk protein concentrate that is produced by passing milk under pressure through a thin, porous membrane to separate the components of milk according to size. Specifically, ultrafiltration allows the smaller lactose, water, mineral, and vitamin molecules to pass through the membrane, while the larger protein and fat molecules (key components for making cheese) are retained and concentrated.
  • Preferred sources are MPC, MPI and MCI, because of the increased protein concentrations therein. Within this subgroup, the preferred starting material is MCI, given its reduced whey protein concentration. All of the above are commercially available.
  • the acidified mixture is cooled at a temperature between 0 °C and 15°C, preferably between 2 and 12 °C, more preferably between 2 and 9 °C.
  • P reduction was improved when cooling from 20 °C to lower temperatures, preferably with the aforementioned ranges.
  • Commercially extensive cooling on industrial scale is not preferred, and the skilled person can optimize the balance between the P reduction and the need for excessive amounts of resources needed for cooling to achieve suitably reduced P levels.
  • Total phosphorous concentrations in the context of the invention can be measured using any conventional method in the art, such as inductively coupled plasma optical emission spectroscopy (ICP-OES) as was used in the examples.
  • ICP-OES inductively coupled plasma optical emission spectroscopy
  • gluconate and/or maleate preferably at least gluconate, is added to the cooled mixture and, it is washed to remove phosphorus.
  • the amount of gluconate and/or maleate is preferably added in an amount of 0.05 - 0.5 g per g micellar casein, preferably 0.1 - 0.4 g per g micellar casein. It is preferred that gluconate is added in an amount of 0.05 - 0.5 g per g micellar casein, preferably 0.1 - 0.4 g per g micellar casein.
  • Casein micelles are present in milk as polydisperse spherical complexes with an average diameter of 200 nm. Casein micelles are heterogeneous, hydrated, dynamic structures with a loose packing and a high porosity. They consist of different types, namely a s i-, a S 2-, P-, and K-casein, and colloidal calcium phosphate (CCP). CCP is essential for maintaining the micellar structure: casein micelles dissociate when CCP is chelated or solubilized. Micelle dissociation can be induced by high-pressure treatment, pH decrease, or calcium chelators. Casein micelles contain two types of phosphorus, which are organic and inorganic phosphorus.
  • the phosphate which is esterified to the casein molecule via a hydroxyl group of serine amino acid is generally called organic phosphate at around 23% in milk, and the phosphate which is associated with the casein molecules in the form of calcium phosphate nanoclusters called as inorganic phosphate at around 32% in milk.
  • organic phosphate the phosphate which is associated with the casein molecules in the form of calcium phosphate nanoclusters
  • inorganic phosphate at around 32% in milk.
  • phosphorus molecules present in the serum phase such as inorganic dissolved phosphate and organic phosphate in the form of esters.
  • the present invention aims to reduce the phosphorus content of skim milk, UF or MCI and a high protein liquid shelf-stable and heat-sterilized enteral nutritional composition by removal of organic phosphate esterified on the serine amino acids and inorganic phosphate present in the calcium phosphate nanoclusters.
  • reducing phosphorus content is challenging because casein micelles can be dissociate and destroy the micellar structure.
  • micellar casein isolate MCI
  • the gluconate and maleate salts do not only enhance the depletion of phosphorus from the casein micelles but also help to maintain the intrinsic structure of the casein micelles. Furthermore, unlike the other chelators mentioned, the addition of the gluconate or maleate salts do not affect the heat stability.
  • maleate disodium maleate is preferred.
  • hydrogen maleate salt such as potassium hydrogen maleate and sodium hydrogen maleate are disclaimed.
  • Gluconate salts are the most preferred chelators, preferably sodium or potassium gluconate.
  • the acidification step is done in order to increase the solubility of phosphorus in casein micelles. It is an important step because adjusting the pH has effect on the particle size, viscosity and stability of the casein micelles.
  • Figure 1 shows that at pH between 6.7 and 6.0 significant reduction of negative charge is observed, and viscosity and particle size remains the same. At pH between 6.0 and 5.6, no significant reduction of negative charge is observed but significant particle size increase is observed. This is attributed to gel formation. At pH 5.4, significant particle size and viscosity increase is observed due to complete collapse of K-casein. Therefore, during the acidification step, the pH is preferably between 6.0 and 7.0, more preferably between 6.0 and 6.7.
  • the present invention further provides dephosphorylated skim milk, ultra-filtered milk (UF milk), including milk protein concentrate (MPC) and milk protein isolate (MPI), or micellar casein isolate (MCI) with gluconate and/or maleate comprises 20 % to 40 % less total phosphorus compared to the untreated corresponding counterpart which can further be characterized by a lack of gluconate and/or maleate, and a phosphorous concentration (i.e. the sum of organic and inorganic phosphorous) which is less than 192 mg/100 ml, in accordance with FSMP regulations.
  • UF milk ultra-filtered milk
  • MCI milk protein isolate
  • MCI micellar casein isolate
  • the total amount of phosphorous in the high-protein composition is at least 10% below the FSMP maximum (i.e. below 72 mg/100 kcal), preferably at least 20% below the FSMP maximum (i.e. below 64 mg/100 kcal).
  • Table 1 below. In accordance, most preferably a reduction between 30 and 40% of the original phosphorous concentration is desired (Table 1).
  • the desired reduction in total phosphorous levels compared to FSMP presented in table 1 can be achieved according to the invention by reducing the amount of phosphorous in a typical micellar casein-providing source with 300 mg P/lOOml by preferably at least 23%, more preferably at least 31%, most preferably at least 38% compared to the original 300 mg P/lOOml concentration in the starting material.
  • total phosphorous levels can be reduced below the FSMP maximum limit, and preferably to less than 90% or even less than 80% of the FSMP-set maximum total phosphorous content, respectively.
  • Table 1 Preferred maximum phosphorus content of high-protein liquid, heat-sterilized enteral nutritional compositions in comparison to commercial products, and FSMP
  • the present application provides a high-protein liquid heat-sterilized enteral nutritional composition with reduced phosphorus content which complies with the requirements of FSMP.
  • the composition comprises phosphorus in an amount between 32 - 78 mg/lOOkcal, preferably 36 - 76 mg/lOOkcal, even more preferably 38 - 74 mg/lOOkcal, most preferably 40 - 70 mg/lOOkcal.
  • liquid enteral nutritional composition refers to an aqueous composition comprising protein, fat and carbohydrates which is to be administered by mouth or by other means, generally by tube feeding, to the stomach or intestines of a patient. Oral administration is preferred.
  • the viscosity is preferably below 500 cP, more preferably below 400 cP, most preferably below 300 cP, as measured at a shear rate of 100 s-1 at 20 °C using a rotational viscosity meter using a cone/plate geometry.
  • the high protein liquid heat-sterilized enteral nutritional composition according to the invention is designed to either supplement a person's diet or to provide complete nutritional support.
  • the composition according to the invention further comprises fat and carbohydrates and preferably a source of vitamins and minerals and/or a source of prebiotics.
  • the composition according to the invention is a nutritionally complete composition.
  • the high-protein composition of the invention is a packaged product ready for transport and marketing. In a preferred embodiment, it is a ready-to-use composition. It is heat-sterilized, and preferably shelf-stable.
  • shelf-stable refers to storage stability.
  • a nutritional composition is shelf-stable if it is storage stable at ambient temperature with respect to microbiological spoilage and physical defects like creaming, gelation, precipitation, etc., for a certain amount of time.
  • the nutritional composition has a shelf-stability of at least one month, more preferably at least 3 months, even more preferably at least 6 months and most preferably at least 12 months after packaging, when stored in a sealed packaging at ambient temperature (20 °C).
  • the invention is not limited to specific sterilization conditions, and in fact the invention renders it possible to subject the high-protein to heat sterilization conditions which are common practice in the field, and which belong to the skilled person's common general knowledge. However, in the field a significant part of the problem of achieving high protein compositions rests in the need for such heat treatment in order to reduce the microbial load to levels that the product can be shelved for extended periods.
  • the total protein preferably provides 16 % to 32 % of the total energy content of the composition, more preferably 18 % to 30 en%, even more preferably 20 % to 28 en%.
  • each of these numbers are in combination with a caloric content of 2.2 - 2.6 kcal/ml.
  • the amount of protein in the composition is preferably 8.8 - 20. 8 g/100 ml, more preferably 9.9 - 19.5 g/100 ml, most preferably 11 - 18.2 g/100 ml.
  • the amount of protein is between 12 and 18 g/100 ml. It is at these higher protein concentrations that the phosphorylated protein i.e. reduced protein concentrations per gram protein, provides an advantage over commercially available high-protein compositions (which cannot meet the FSMP guidelines when using profound amounts of MC).
  • micellar casein preferably 70 - 95 wt%, more preferably 75 - 95 wt%, most preferably 80 - 95 wt% of all proteinaceous matter.
  • the term 'micellar casein' refers to the structure of casein proteins in milk which is commonly known in the art and comprises of the different casein proteins aSl, aS2, P, K casein.
  • the size of the micelles can vary. In unprocessed milk the micelles can vary between about 100 and 500 nm, but after dephosphorylation using the method of the invention, the size of the micelles can even vary wider, yet the micellar structure is essentially retained.
  • compositions according to the invention further comprises at least 2 mineral levels according to the ranges according to the table below, more preferably at least 3, even more preferably at least 4, most preferably all mineral levels for Na, K, Cl, Ca and Mg according to the FSMP recommended levels requirements summarized in according to the table below (which is an extract of table 2 of the above EU Directive):
  • the composition comprises 30 - 175 mg sodium per 100 kcal of the composition.
  • the composition preferably comprises sodium in an amount between 30 - 140 mg/lOOkcal, more preferably 32 - 100 mg/lOOkcal, even more preferably 34 - 80 mg/lOOkcal.
  • the composition comprises from 70 - 250 mg sodium per 100ml of the liquid composition, preferably 80 - 230 mg sodium per 100ml of the liquid composition.
  • the composition preferably comprises potassium.
  • the composition comprises 80 - 295 mg potassium per 100 kcal of the composition.
  • the composition comprises potassium in an amount between 85 - 250 mg/lOOkcal, more preferably 90 - 200 mg/lOOkcal, even more preferably 95 - 150 mg/lOOkcal.
  • the composition preferably comprises from 180 - 400 mg potassium per 100ml of the liquid composition, more preferably 200 - 380 mg potassium per 100ml of the liquid composition.
  • potassium is not provided in the form of a potassium phosphate salt.
  • a suitable form could be potassium gluconate and/or dipotassium maleate.
  • the process of dephosphorylation does not affect nor is it affected by the presence of calcium in the composition.
  • calcium concentrations in the skim milk, UF milk or MCI are about 30 mg per gram protein. It is preferred to maintain Ca levels within FSMP standards, i.e. between 35 and 175 mg Ca per 100 kcal.
  • the composition preferably comprises chlorine (Cl) in an amount between 30 - 175 mg/lOOkcal, preferably 35 - 150 mg/lOOkcal, even more preferably 40 - 140 mg/lOOkcal.
  • the composition comprises from 100 - 380 mg chlorine per 100ml of the liquid composition, preferably 110 - 350 mg chlorine per 100ml of the liquid composition.
  • a liquid shelf-stable/heat-sterilized enteral nutritional composition comprising 12 - 16 g/100 ml protein and 2.0 - 3.0 kcal/ml caloric content, preferably 2.2 - 2.6 kcal/ml, wherein the protein comprises micellar casein (MC), whey protein (WP) and optionally caseinate (CAS), wherein there is at least 70wt% MC and less than 15 wt% WP, based on total protein content, the composition preferably comprising 7 - 12 g/100 ml fat and 18 - 30 g/100 ml digestible carbohydrates, and wherein the composition has a total amount of phosphorous less than 192 mg/100 ml and/or 30 - 80 mg/lOOkcal, preferably at least 30 - 80 mg/lOOkcal, more preferably less than 192 mg/100 ml and 30 - 80 mg/lOOkcal.
  • MC micellar casein
  • WP whey protein
  • WP when WP is included in the composition in the invention, this is preferably controlled to reduced amounts i.e. less than 15 wt%, most preferably less than 10 wt%, based on total protein.
  • WP can also be provided in intact and/or hydrolyzed form. The above is the sum of both hydrolyzed and intact WP.
  • a measure for the extent of hydrolysation of the whey protein is the "degree of hydrolysation" (DH).
  • the DH is defined as the percentage of the total number of peptide bonds in a protein that has been cleaved during hydrolysis.
  • the DH of a protein may e.g.
  • the source of whey protein may already comprise a certain (small) amount of peptide fractions, before being subjected to the hydrolysis process.
  • the values for the degree of hydrolysation as described herein are corrected for this presence of peptide-fractions in the whey protein source, in other words, the values for the DH are corrected for the natural DH of whey protein.
  • the DH thus relates to the additional hydrolysation that was obtained via the intentional hydrolysis process.
  • the composition comprises hydrolysed whey protein
  • it preferably has a degree of hydrolysation of 1-25%, preferably in the range of 5 to 25%.
  • the degree of hydrolysation as used herein is corrected for the natural degree of hydrolysation of the whey protein source, i.e. the whey protein that was used for the preparation of the hydrolysed whey protein.
  • the composition may also comprise limited amounts of caseinate.
  • Most MCI sources have WP contents below 10%, preferably below 5 wt%, based on total protein.
  • a combination of micellar casein and caseinate is particularly preferred when the micellar casein is provided by a source which comprises more than 10 wt% whey protein, such as MPC or MPI (with a weight ratio of micellar casein to whey protein of 80:20).
  • the caseinates may be added to the micellar casein in order to control (reduce) the amount of whey protein levels in the protein fraction as described above.
  • the weight ratio of micellar casein to caseinate ranges from 90:10 to 60:40.
  • Na-caseinate, Mg-caseinate, K-caseinate, Ca-caseinate or any mixture thereof or combinations thereof such as Na/K-caseinate and Na/Mg caseinate are used as the source of caseinate.
  • Ca-caseinate, or a caseinate comprising Ca is not used, as the micellar casein already contains a sufficient amount of calcium.
  • Na/K caseinates provide better taste.
  • composition preferably comprises: 16-35 en% protein
  • en% refers to % of total energy of the composition. It thus refers to energy percentage representing the relative amount that a constituent contributes to the total caloric value of the composition.
  • the amounts of energy provided by proteins, fats and carbohydrates can be estimated e.g. using Atwater factor wherein: 4 kcal per gram (kcal/g) (17 kJ/g) for protein (and amino acids), 4 kcal/g for digestible carbohydrates and 9 kcal/g (37 kJ/g) for fat.
  • the liquid composition preferably comprises 7 - 12 g/100 ml fat and 18 - 30 g/100 ml digestible carbohydrates.
  • the liquid nutritional composition according to the invention preferably comprises fat, said fat providing between 30 to 55 % of the total energy content of the composition.
  • the compositions according to the invention comprise vegetable fats, preferably rapeseed oil, sunflower oil, corn oil, soybean oil, canola oil, or combinations thereof.
  • the fat may include medium chain triglycerides (MCT, mainly 8 to 10 carbon atoms long), long chain triglycerides (LCT) or any combination of the two types, according to the desired benefits.
  • MCT medium chain triglycerides
  • the fat comprises 30 to 60 wt% of animal or algal fat, 40 to 70 wt% of vegetable fat and optionally 0 to 20 wt% of MCTs based on total fat of the composition.
  • the animal fat preferably comprises a low amount of milk fat, i.e. lower than 6 wt%, especially lower than 3 wt%.
  • a mixture of corn oil, egg oil, and/or canola oil and specific amounts of marine oil are used.
  • Egg oils, fish oils and algal oils are a preferred source of non-vegetable fats.
  • compositions that are to be consumed orally in order to prevent formation of off- flavours and to decrease a fishy after-taste, it is recommended to select ingredients that are relatively low in docosahexanoic acid (DHA), i.e. less than 6 wt%, preferably less than 4 wt% of the fat.
  • DHA docosahexanoic acid
  • Marine oils containing DHA are preferably present in the composition according to the invention in an amount lower than 25 wt%, preferably lower than 15 wt% of the fat.
  • inclusion of eicosapentanoic acid (EPA) is highly desirable for obtaining the maximum health effect.
  • the amount of EPA ranges preferably between 4 wt% and 15 wt%, more preferably between 8 wt% and 13 wt% of the fat.
  • the weight ratio EPA:DHA is advantageously at least 6:4, for example between 2:1 1 and 10:1.
  • the liquid nutritional composition according to the invention may beneficially comprise an emulsifier.
  • an emulsifier may be used, such as lecithin, and generally the emulsifier contributes to the energy content of the fat in said composition.
  • the liquid nutritional composition according to the invention comprises digestible carbohydrates, said carbohydrates providing between 30 to 55 % of the total energy content of the composition.
  • Suitable digestible carbohydrates are glucose, fructose, sucrose, lactose, trehalose, palatinose, corn syrup, malt, maltose, isomaltose, partially hydrolysed corn starch, maltodextrins, glucose oligo- and poly-saccharides.
  • the digestible carbohydrates include trehalose or isomaltulose.
  • Trehalose/isomaltulose can reduce sweetness compared to sucrose, because the relative sweetness is as sucrose (100), trehalose (45), isomaltulose (40-50).
  • the viscosity as measured is similar whichever of the three is used for a digestible carbohydrate.
  • panelists favored trehalose over sucrose in the high-protein composition of the invention, for it was perceived less viscous.
  • trehalose is a preferred choice of carbohydrate, as it gives rise to a low (perceived) viscosity, no undesired Maillard reactions and it has a sweetness about half of that of sucrose.
  • the digestible carbohydrates include trehalose in an amount of 20 to 60 wt% of the digestible carbohydrates, more preferably in an amount of 20 % to 45 wt%, even more preferably in an amount of 25 to 45 wt% of the digestible carbohydrates; in a most preferred embodiment, the remainder is provided by maltodextrins with DE 16-20.
  • the present invention further refers to the use of the compositions described herein for preventing or treating malnutrition in a person in need thereof, preferably a malnourished person and/or elderly preferably at least 50 years of age.
  • the invention also concerns a (non-therapeutic) method of providing nutrition to a person in need thereof, preferably a malnourished person and/or elderly preferably at least 50 years of age, the method comprising enterally, preferably orally, administering the liquid nutritional composition according to the invention.
  • Example 1 - MCI was tested using different types of calcium chelators (Table 2). pH and T were also varied.
  • micellar casein isolate solution was prepared in demineralized water at 60°C, 105 grams (7% (w/v)) of powdered MCI-88 from Friesland Campina was dissolved. To dissolve the powdered MCI, the solution was placed in a pre-heated (60°C) water bath (Julabo GmbH, Boven-Leeuwen, The Netherlands) where the solution was stirred at 1000 rpm using an overhead stirrer (IKA, Staufen, Germany). When a complete powder dispersion was observed, the dispersion was left in the water bath of 60°C for 1 hour while continuously stirred at 200 rpm.
  • the dispersion was homogenized with the ultra-turrax (IKA, Oude Vijvers, the Netherlands) for 5 minutes at 25.000 rpm. Then, the protein dispersion was further homogenized using a two-staged GEA Pony NS2006L pilot-scale homogenizer (Parma, Italy) with a first valve pressure of 300 bar and a second valve pressure of 50 bar. To confirm a complete dissolution of the particles in the solution, the particle size distribution was measured with the Mastersizer 3000 (Malvern Analytical Ltd., Malvern, United Kingdom). In addition, the dry matter was measured by CEM moisture analyser (Mettler Toledo, Tiel, the Netherlands). The sample was stored overnight in de fridge at 4°C to equilibrate.
  • the MCI solution was acidified to pH 6.0 ( ⁇ 0.05), and chelator with a concentration of 70 mEq K/L was added.
  • the chelator was selected from K-gluconate, disodium maleate and K-citrate.
  • Microfiltration and/or diafiltration was performed and the retentate was obtained.
  • the amount of phosphorous in starting material and in the retentate was measured using Inductively Coupled Plasma Optical Emission spectroscopy (ICP-OES).
  • ICP-OES Inductively Coupled Plasma Optical Emission spectroscopy
  • the amount of P in the starting material was 17.6 - 20 mg.
  • the particle size distribution was measured using the Mastersizer 3000 (Malvern Analytical Ltd., Malvern, United Kingdom). The data was processed using the Mastersizer 3000 Software v3.80 (Malvern Analytica Ltd., Malvern, United Kingdom). All measurements were performed three times.
  • the Zeta-potential was measured with the Zetasizer Nano Z (Malvern Analytical Ltd., Malvern, United Kingdom) equipped with 4 mW He-Ne laser. Disposable folded capillary Zetasizer Nano cells of 1.5 mL (DTS1060, Malvern Instruments, Worcestershire, UK) were used for the measurements. The samples were 100 times diluted in the supernatant of the ultracentrifugated samples. Analyses were performed at ambient cell temperature and a voltage of 100 V. Data were processed with Zetasizer software (Malvern Analytical Ltd., Malvern, United Kingdom).
  • Viscosity was measured at a shear rate of 100 s-1 at 20 °C using a rotational viscosity meter using a cone/plate geometry.
  • composition A Composition A
  • dephosphorylated MCI-88 (retentate) (prepared using 70 mEq potassium gluconate with pH 6.0 and 4 °C cooling) was used as starting material.
  • the retentate was stirred using a magnetic stirrer and pre-warmed to 55°C with a hotplate stirrer (imLab IKA plate, Oude Vijvers, The Netherlands).
  • sodium caseinate and sugar, both in powder form were mixed before they were added to the retentate. When dissolved, the maltodextrin was added. Before adding citrate and calcium chloride, the minerals were pre-mixed in water in a 1:10 ratio.
  • magnesium and citric acid were premixed and subsequently added to the mixture while stirring.
  • the left-over minerals were added one at the time directly to the mixture until dissolved.
  • the oil mixture of canola oil and lecithin was first blended and preheated till 65°C. Then, the oil mixture was added and homogenised using the ultra-turrax (IKA, Oude Vijvers, the Netherlands) for 5 minutes at 25,000 rpm to prevent phase separation. Each sample (retentate and total product) was transferred to pressure-resistant DURAN culture tubes to prevent the product from boiling.
  • the samples were put into a temperature-controlled oil bath (Julabo GmbH, Boven-Leeuwen, the Netherlands) and heated at 127°C. During the heat treatment, the samples were turned upside down. After 5, 10 and 20 minutes of heating the heat coagulation time (HCT) was measured via visual inspection. After 20 minutes the samples were cooled with icepacks to 5 °C.
  • HCT heat coagulation time
  • micellar casein Treatment with gluconate resulted in the most profound P reduction in the micellar casein, and the micellar casein structure was retained, particularly stable in case of gluconate.
  • FIG 1 there is plotted the zeta-potential (A), particle size (B) and viscosity (C) of a micellar casein isolate when subjected to pH variation and cooling variation. In those experiments no chelator was applied.
  • Figures 1A and IB both clearly show that below pH 6 the micellar structure is lost, even though this may not be directly observed in the viscosity profile in Figure 1C. pH values below 6.0 resulted in (partly) aggregated casein micelles. Moreover, it was suggested that internal casein micelle damaged, in the form of casein molecule solubilization (especially -casein), which increased at pH 6.0 and progressed with a reducing pH.
  • Example 2A liquid heat-sterilized enteral nutritional composition
  • total P is 165 mg/100 ml or 68.7 mg P/lOOkcal.
  • Example 2B liquid heat-sterilized enteral nutritional composition
  • the recipe of example 2A has the following target mineral levels, all at least 20% less than the maximum values set by FSMP. These target numbers are provided together with the FSMP ranges, and the observed amounts in corresponding high-protein compositions high in micellar casein currently marketed.
  • Table 6 shows an overview of different minerals that had been tested in an attempt to replace monosodium phosphate in the process of reconstitution of casein micelles, seeking a way to reduce phosphorous while retaining the desired micellar casein structure necessary to prepare high-protein compositions.

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Abstract

L'invention concerne un procédé de déphosphorylation de lait écrémé, de lait ultra-filtré (lait UF) ou d'isolat de caséine micellaire (MCI) consistant à (i) acidifier un lait écrémé, un lait UF ou un MCI à un pH de préférence non inférieur à 6,0, de préférence entre 6,0 et 6,7, (ii) refroidir le lait écrémé, le lait UF ou le MCI acidifié à une température comprise entre 0°C et 15°C, (iii) ajouter du gluconate et/ou de maléate au lait écrémé, au lait UF ou au MCI refroidi et, (iv) extraire par lavage le phosphore du lait écrémé, du lait UF ou du MCI, déphosphorylant ainsi le lait écrémé, le lait UF ou le MCI, de préférence dans une mesure telle que la teneur en phosphore totale du lait écrémé, du lait UF ou du MCI soit réduite d'au moins 20%, de préférence de 20 à 40%, plus préférablement de 30 à 40% par rapport à la substance obtenue à l'étape (i). L'invention concerne également du lait écrémé, du lait UF ou du MCI déphosphorylé pouvant être obtenu par le procédé de l'invention, et une composition nutritionnelle entérale thermostérilisée liquide contenant 2,0 à 3,0 kcal/ml. 16 à 35% en énergie sont fourni par la protéine, la combinaison de la teneur calorique et de la teneur calorique relative des protéines est choisie de telle sorte qu'il y ait 10 à 18 g/100 ml de protéine, de préférence 12 à 18 g/100 ml de protéine dans la composition. La protéine contient de la caséine micellaire (MC), de la protéine de lactosérum (WP) et éventuellement du caséinate (CAS). Il y a au moins 70% en poids de MC et moins de 15% en poids de WP. La composition a une quantité totale de phosphore inférieure à 192 mg/100 ml et/ou 30 à 80 mg/100 kcal, de préférence au moins 30 à 80 mg/100 kcal, idéalement moins de 192 mg/100 ml et 30 à 80 mg/100 kcal.
PCT/EP2023/061832 2023-05-04 2023-05-04 Déphosphorylation de lait écrémé, de lait ultra-filtré ou d'isolat de caséine micellaire WO2024227514A1 (fr)

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PCT/EP2024/062051 WO2024227851A1 (fr) 2023-05-04 2024-05-02 Déphosphorylation de lait écrémé, de lait ultrafiltré ou d'isolat de caséine micellaire

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WO2009072885A1 (fr) 2007-12-05 2009-06-11 N.V. Nutricia Composition nutritionnelle entérale liquide riche en énergie
US20110159163A1 (en) * 2008-07-23 2011-06-30 N.V. Nutricia Method for filtering milk
WO2013129925A1 (fr) 2012-02-28 2013-09-06 N.V. Nutricia Composition nutritionnelle liquide énergétique ayant des propriétés organoleptiques améliorées
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